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Molecular cytogenetic mapping of chromosomal fragments and immunostaining of kinetochore proteins in Beta. β染色体片段的分子细胞遗传学定位和着丝点蛋白的免疫染色。
Pub Date : 2009-01-01 Epub Date: 2009-11-08 DOI: 10.1155/2009/721091
Daryna Dechyeva, Thomas Schmidt

By comparative multicolor FISH, we have physically mapped small chromosome fragments in the sugar beet addition lines PRO1 and PAT2 and analyzed the distribution of repetitive DNA families in species of the section Procumbentes of the genus Beta. Six repetitive probes were applied, including genotype-specific probes-satellites pTS4.1, pTS5, and pRp34 and a dispersed repeat pAp4, the telomere (TTTAGGG)(n), and the conserved 18S-5.8S-25S rRNA genes. Pachytene-FISH analysis of the native centromere organization allowed proposing the origin of PRO1 and PAT2 fragments. Comparative analysis of the repetitive DNA distribution and organization in the wild beet and in the addition lines allowed the development of a physical model of the chromosomal fragments. Immunostaining revealed that the PRO1 chromosome fragment binds alpha-tubulin and the serine 10-phosphorylated histone H3 specific for the active centromere. This is the first experimental detection of the kinetochore proteins in Beta showing their active involvement in chromosome segregation in mitosis.

通过比较多色FISH,我们对甜菜加成系PRO1和PAT2的小染色体片段进行了物理定位,并分析了重复DNA家族在β属Procumbentes部分种中的分布。使用了6个重复探针,包括基因型特异性探针-卫星pTS4.1、pTS5和pRp34,以及分散重复的pAp4、端粒(TTTAGGG)(n)和保守的18S-5.8S-25S rRNA基因。原生着丝粒组织的Pachytene-FISH分析允许提出PRO1和PAT2片段的起源。通过对野生甜菜和附加品系中重复DNA分布和组织的比较分析,建立了染色体片段的物理模型。免疫染色显示,PRO1染色体片段结合了α -微管蛋白和丝氨酸10-磷酸化的组蛋白H3,这是活性着丝粒所特有的。这是第一次实验检测到β中的着丝点蛋白,表明它们积极参与有丝分裂中的染色体分离。
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引用次数: 4
PROC QTL-A SAS Procedure for Mapping Quantitative Trait Loci. QTL-A SAS定量性状位点定位程序。
Pub Date : 2009-01-01 Epub Date: 2009-12-08 DOI: 10.1155/2009/141234
Zhiqiu Hu, Shizhong Xu

Statistical analysis system (SAS) is the most comprehensive statistical analysis software package in the world. It offers data analysis for almost all experiments under various statistical models. Each analysis is performed using a particular subroutine, called a procedure (PROC). For example, PROC ANOVA performs analysis of variances. PROC QTL is a user-defined SAS procedure for mapping quantitative trait loci (QTL). It allows users to perform QTL mapping for continuous and discrete traits within the SAS platform. Users of PROC QTL are able to take advantage of all existing features offered by the general SAS software, for example, data management and graphical treatment. The current version of PROC QTL can perform QTL mapping for all line crossing experiments using maximum likelihood (ML), least square (LS), iteratively reweighted least square (IRLS), Fisher scoring (FISHER), Bayesian (BAYES), and empirical Bayes (EBAYES) methods.

统计分析系统(SAS)是目前世界上最全面的统计分析软件包。它提供了各种统计模型下几乎所有实验的数据分析。每次分析都是使用特定的子例程执行的,称为过程(PROC)。例如,PROC ANOVA执行方差分析。PROC QTL是一个用户自定义的SAS程序,用于定位数量性状位点(QTL)。它允许用户在SAS平台内执行连续和离散特征的QTL映射。PROC QTL的用户能够利用通用SAS软件提供的所有现有功能,例如,数据管理和图形处理。当前版本的PROC QTL可以使用最大似然(ML)、最小二乘(LS)、迭代加权最小二乘(IRLS)、Fisher评分(Fisher)、贝叶斯(BAYES)和经验贝叶斯(EBAYES)方法对所有的杂交实验进行QTL定位。
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引用次数: 61
Reproducible RNA preparation from sugarcane and citrus for functional genomic applications. 从甘蔗和柑橘中制备用于功能基因组应用的可重复 RNA。
Pub Date : 2009-01-01 Epub Date: 2010-01-27 DOI: 10.1155/2009/765367
Mona B Damaj, Phillip D Beremand, Marco T Buenrostro-Nava, Beth Riedel, Joe J Molina, Siva P Kumpatla, Terry L Thomas, T Erik Mirkov

High-throughput functional genomic procedures depend on the quality of the RNA used. Copurifying molecules can negatively impact the functionality of some plant RNA preparations employed in these procedures. We present a simplified, rapid, and scalable SDS/phenol-based method that provides the high-quantity and -quality RNA required by the newly emerging biotechnology applications. The method is applied to isolating RNA from tissues of two biotechnologically important crop plants, sugarcane and citrus, which provide a challenge due to the presence of fiber, polysaccharides, or secondary metabolites. The RNA isolated by this method is suitable for several downstream applications including northern blot hybridization, microarray analysis, and quantitative RT-PCR. This method has been used in a diverse range of projects ranging from screening plant lines overexpressing mammalian genes to analyzing plant responses to viral infection and defense signaling molecules.

高通量功能基因组程序取决于所用 RNA 的质量。共聚分子会对这些程序中使用的某些植物 RNA 制剂的功能产生负面影响。我们提出了一种基于 SDS/苯酚的简化、快速和可扩展的方法,可提供新兴生物技术应用所需的高数量和高质量的 RNA。该方法适用于从甘蔗和柑橘这两种具有重要生物技术价值的作物组织中分离 RNA。用这种方法分离的 RNA 适用于多种下游应用,包括北印迹杂交、微阵列分析和定量 RT-PCR。这种方法已被用于多种项目,从筛选过表达哺乳动物基因的植物品系到分析植物对病毒感染和防御信号分子的反应。
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引用次数: 0
QTL mapping of a high protein digestibility trait in Sorghum bicolor. 高粱高蛋白质消化率性状的QTL定位。
Pub Date : 2009-01-01 Epub Date: 2009-07-07 DOI: 10.1155/2009/471853
Jennifer A Winn, R Esten Mason, Adriana L Robbins, William L Rooney, Dirk B Hays

Compared with other cereal grains, Sorghum bicolor shows lower protein digestibility. The low digestibility is thought to result from disulfide cross linking in the beta- and gamma-kafirins. In contrast, the single recessive high digestibility/high lysine content (HD) mutation which confers greater grain digestibility exists in sorghum that is thought to result from reduced accumulation of gamma-kafirin that allows greater access to the high digestible alpha-kafarin fraction. In an effort to both clearly define the molecular basis for the HD trait and develop tools to improve the introgression of this difficult-to-screen trait, this study focuses on mapping the QTLs linked to this trait. While the HD trait has been defined as a single recessive gene, our results uncovered that two major QTLs on chromosome 1 are associated with protein digestibility-one QTL (locus 1 from the HD parent) unfavorably affects digestibility and one QTL (locus 2 from the HD parent) only 20 cM away favorably affects digestibility. A contrast analysis between genotypic groups at these two loci shows that a higher level of protein digestibility may be obtained when this linkage in repulsion is broken and favorable alleles are allowed to recombine.

与其他谷物相比,高粱的蛋白质消化率较低。低消化率被认为是由于-和-kafirin中的二硫交联所致。相比之下,单隐性高消化率/高赖氨酸含量(HD)突变使高粱的谷物消化率更高,这被认为是由于γ -卡弗林积累减少,从而更容易获得高消化的α -卡弗林部分。为了清楚地定义HD性状的分子基础,并开发工具来改善这种难以筛选的性状的渗入,本研究的重点是绘制与该性状相关的qtl。虽然HD性状被定义为单隐性基因,但我们的研究结果发现,1号染色体上的两个主要QTL与蛋白质消化率相关——一个QTL(来自HD亲本的1号位点)不利影响消化率,另一个QTL(来自HD亲本的2号位点)仅20 cM远,有利影响消化率。对这两个基因座基因型组的对比分析表明,当这种排斥连锁被打破并允许有利等位基因重组时,可以获得更高水平的蛋白质消化率。
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引用次数: 34
Advances in maize genomics and their value for enhancing genetic gains from breeding. 玉米基因组学的进展及其对提高育种遗传收益的价值。
Pub Date : 2009-01-01 Epub Date: 2009-08-12 DOI: 10.1155/2009/957602
Yunbi Xu, Debra J Skinner, Huixia Wu, Natalia Palacios-Rojas, Jose Luis Araus, Jianbing Yan, Shibin Gao, Marilyn L Warburton, Jonathan H Crouch

Maize is an important crop for food, feed, forage, and fuel across tropical and temperate areas of the world. Diversity studies at genetic, molecular, and functional levels have revealed that, tropical maize germplasm, landraces, and wild relatives harbor a significantly wider range of genetic variation. Among all types of markers, SNP markers are increasingly the marker-of-choice for all genomics applications in maize breeding. Genetic mapping has been developed through conventional linkage mapping and more recently through linkage disequilibrium-based association analyses. Maize genome sequencing, initially focused on gene-rich regions, now aims for the availability of complete genome sequence. Conventional insertion mutation-based cloning has been complemented recently by EST- and map-based cloning. Transgenics and nutritional genomics are rapidly advancing fields targeting important agronomic traits including pest resistance and grain quality. Substantial advances have been made in methodologies for genomics-assisted breeding, enhancing progress in yield as well as abiotic and biotic stress resistances. Various genomic databases and informatics tools have been developed, among which MaizeGDB is the most developed and widely used by the maize research community. In the future, more emphasis should be given to the development of tools and strategic germplasm resources for more effective molecular breeding of tropical maize products.

玉米是世界热带和温带地区重要的粮食、饲料、饲草和燃料作物。遗传、分子和功能层面的多样性研究表明,热带玉米种质、陆生品系和野生近缘种蕴藏着更广泛的遗传变异。在所有类型的标记中,SNP 标记日益成为玉米育种中所有基因组学应用的首选标记。基因图谱绘制是通过传统的连锁图谱绘制发展起来的,最近则是通过基于连锁不平衡的关联分析发展起来的。玉米基因组测序最初侧重于基因丰富的区域,现在的目标是提供完整的基因组序列。传统的基于插入突变的克隆最近得到了基于EST和图谱的克隆的补充。转基因和营养基因组学正在迅速推进以抗虫性和谷物品质等重要农艺性状为目标的领域。基因组学辅助育种方法取得了长足进步,提高了产量以及抗非生物性和生物性胁迫的能力。已开发出各种基因组数据库和信息学工具,其中 MaizeGDB 是最先进的,并被玉米研究界广泛使用。今后,应更加重视开发工具和战略种质资源,以更有效地进行热带玉米产品的分子育种。
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引用次数: 0
Transcriptomic analysis of starch biosynthesis in the developing grain of hexaploid wheat. 六倍体小麦籽粒发育中淀粉生物合成的转录组学分析。
Pub Date : 2009-01-01 Epub Date: 2010-03-08 DOI: 10.1155/2009/407426
Boryana S Stamova, Debbie Laudencia-Chingcuanco, Diane M Beckles

The expression of genes involved in starch synthesis in wheat was analyzed together with the accumulation profiles of soluble sugars, starch, protein, and starch granule distribution in developing caryopses obtained from the same biological materials used for profiling of gene expression using DNA microarrays. Multiple expression patterns were detected for the different starch biosynthetic gene isoforms, suggesting their relative importance through caryopsis development. Members of the ADP-glucose pyrophosphorylase, starch synthase, starch branching enzyme, and sucrose synthase gene families showed different expression profiles; expression of some members of these gene families coincided with a period of high accumulation of starch while others did not. A biphasic pattern was observed in the rates of starch and protein accumulation which paralleled changes in global gene expression. Metabolic and regulatory genes that show a pattern of expression similar to starch accumulation and granule size distribution were identified, suggesting their coinvolvement in these biological processes.

利用DNA芯片分析了小麦淀粉合成相关基因的表达、可溶性糖、淀粉、蛋白质和淀粉颗粒在发育颖果中的积累谱,并分析了颖果中淀粉合成相关基因的表达谱。不同淀粉生物合成基因异构体的多种表达模式被检测到,表明它们在颖果发育中的相对重要性。adp -葡萄糖焦磷酸化酶、淀粉合酶、淀粉分支酶和蔗糖合酶基因家族的成员表现出不同的表达谱;这些基因家族的一些成员的表达与淀粉的高积累时期一致,而其他成员则没有。淀粉和蛋白质的积累速率呈双相模式,这与全球基因表达的变化相平行。代谢和调控基因表现出类似于淀粉积累和颗粒大小分布的表达模式,表明它们共同参与这些生物过程。
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引用次数: 40
Heterelogous expression of plant genes. 植物基因的异源表达。
Pub Date : 2009-01-01 Epub Date: 2009-08-06 DOI: 10.1155/2009/296482
Filiz Yesilirmak, Zehra Sayers

Heterologous expression allows the production of plant proteins in an organism which is simpler than the natural source. This technology is widely used for large-scale purification of plant proteins from microorganisms for biochemical and biophysical analyses. Additionally expression in well-defined model organisms provides insights into the functions of proteins in complex pathways. The present review gives an overview of recombinant plant protein production methods using bacteria, yeast, insect cells, and Xenopus laevis oocytes and discusses the advantages of each system for functional studies and protein characterization.

异源表达允许在生物体中产生比天然来源更简单的植物蛋白。该技术广泛应用于微生物中植物蛋白的大规模纯化,用于生物化学和生物物理分析。此外,在定义明确的模式生物中的表达提供了对复杂途径中蛋白质功能的见解。本文综述了利用细菌、酵母、昆虫细胞和非洲爪蟾卵母细胞生产重组植物蛋白的方法,并讨论了每种系统在功能研究和蛋白质表征方面的优势。
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引用次数: 46
A Nest of LTR Retrotransposons Adjacent the Disease Resistance-Priming Gene NPR1 in Beta vulgaris L. U.S. Hybrid H20. 甜菜杂种H20抗病启动基因NPR1附近的LTR反转录转座子巢
Pub Date : 2009-01-01 Epub Date: 2009-04-15 DOI: 10.1155/2009/576742
David Kuykendall, Jonathan Shao, Kenneth Trimmer

A nest of long terminal repeat (LTR) retrotransposons (RTRs), discovered by LTR_STRUC analysis, is near core genes encoding the NPR1 disease resistance-activating factor and a heat-shock-factor-(HSF-) like protein in sugarbeet hybrid US H20. SCHULTE, a 10 833 bp LTR retrotransposon, with 1372 bp LTRs that are 0.7% divergent, has two ORFs with unexpected introns but encoding a reverse transcriptase with rve and Rvt2 domains similar to Ty1/copia-type retrotransposons and a hypothetical protein. SCHULTE produced significant nucleotide BLAST alignments with repeat DNA elements from all four families of plants represented in the TIGR plant repeat database (PRD); the best nucleotide sequence alignment was to ToRTL1 in Lycopersicon esculentum. A second sugarbeet LTR retrotransposon, SCHMIDT, 11 565 bp in length, has 2561 bp LTRs that share 100% identity with each other and share 98-99% nucleotide sequence identity over 10% of their length with DRVs, a family of highly repetitive, relatively small DNA sequences that are widely dispersed over the sugarbeet genome. SCHMIDT encodes a complete gypsy-like polyprotein in a single ORF. Analysis using LTR_STRUC of an in silico deletion of both of the above two LTR retrotransposons found that SCHULTE and SCHMIDT had inserted within an older LTR retrotransposon, resulting in a nest that is only about 10 Kb upstream of NPR1 in sugarbeet hybrid US H20.

通过LTR_STRUC分析,在甜菜杂交品种US H20中编码NPR1抗病激活因子和HSF-样蛋白的核心基因附近发现了一个长末端重复(LTR)反转录转座子(RTRs)巢。SCHULTE是一个10833 bp的LTR逆转录转座子,其1372 bp的LTRs有0.7%的差异,具有两个意想不到的内含子的orf,但编码具有与Ty1/复制型逆转录转座子相似的rve和Rvt2结构域的逆转录酶和一种假设的蛋白质。SCHULTE与TIGR植物重复序列数据库(PRD)中所有四个植物家族的重复DNA元素进行了显著的核苷酸BLAST比对;番茄中ToRTL1的核苷酸序列比对效果最好。另一个甜菜LTR反转录转座子SCHMIDT长度为11 565 bp,其长度为2561 bp的LTR彼此具有100%的同源性,并且与drv具有超过10%长度的98-99%的核苷酸序列同源性,drv是一个高度重复的相对较小的DNA序列家族,广泛分布在甜菜基因组中。施密特在一个ORF中编码了一个完整的吉普赛样多蛋白。利用LTR_STRUC对上述两个LTR反转录转座子的缺失进行分析发现,SCHULTE和SCHMIDT插入了一个较老的LTR反转录转座子,在甜菜杂交株US H20中,在NPR1上游约10 Kb处形成了一个巢。
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引用次数: 19
Analysis of Gene Expression Profiles in Leaf Tissues of Cultivated Peanuts and Development of EST-SSR Markers and Gene Discovery. 分析栽培花生叶组织的基因表达谱并开发 EST-SSR 标记和基因发现。
Pub Date : 2009-01-01 Epub Date: 2009-06-24 DOI: 10.1155/2009/715605
Baozhu Guo, Xiaoping Chen, Yanbin Hong, Xuanqiang Liang, Phat Dang, Tim Brenneman, Corley Holbrook, Albert Culbreath

Peanut is vulnerable to a range of foliar diseases such as spotted wilt caused by Tomato spotted wilt virus (TSWV), early (Cercospora arachidicola) and late (Cercosporidium personatum) leaf spots, southern stem rot (Sclerotium rolfsii), and sclerotinia blight (Sclerotinia minor). In this study, we report the generation of 17,376 peanut expressed sequence tags (ESTs) from leaf tissues of a peanut cultivar (Tifrunner, resistant to TSWV and leaf spots) and a breeding line (GT-C20, susceptible to TSWV and leaf spots). After trimming vector and discarding low quality sequences, a total of 14,432 high-quality ESTs were selected for further analysis and deposition to GenBank. Sequence clustering resulted in 6,888 unique ESTs composed of 1,703 tentative consensus (TCs) sequences and 5185 singletons. A large number of ESTs (5717) representing genes of unknown functions were also identified. Among the unique sequences, there were 856 EST-SSRs identified. A total of 290 new EST-based SSR markers were developed and examined for amplification and polymorphism in cultivated peanut and wild species. Resequencing information of selected amplified alleles revealed that allelic diversity could be attributed mainly to differences in repeat type and length in the SSR regions. In addition, a few additional INDEL mutations and substitutions were observed in the regions flanking the microsatellite regions. In addition, some defense-related transcripts were also identified, such as putative oxalate oxidase (EU024476) and NBS-LRR domains. EST data in this study have provided a new source of information for gene discovery and development of SSR markers in cultivated peanut. A total of 16931 ESTs have been deposited to the NCBI GenBank database with accession numbers ES751523 to ES768453.

花生易受一系列叶面病害的侵袭,如番茄斑萎病毒(TSWV)引起的斑萎病、早期叶斑病(Cercospora arachidicola)和晚期叶斑病(Cercosporidium personatum)、南方茎腐病(Sclerotium rolfsii)和枯萎病(Sclerotinia minor)。在这项研究中,我们报告了从一个花生栽培品种(Tifrunner,抗 TSWV 和叶斑病)和一个育种品系(GT-C20,易感 TSWV 和叶斑病)的叶组织中生成的 17,376 个花生表达序列标签(ESTs)。在修剪载体和剔除低质量序列后,共挑选出 14,432 个高质量 EST 进行进一步分析,并将其存入 GenBank。通过序列聚类,得到了 6,888 个独特的ESTs,其中包括 1,703 个暂定共识(TC)序列和 5185 个单体序列。此外,还发现了大量代表未知功能基因的ESTs(5717个)。在这些独特序列中,共鉴定出 856 个 EST-SSR。共开发了 290 个基于 EST 的新 SSR 标记,并对其在栽培花生和野生物种中的扩增和多态性进行了检验。对所选扩增等位基因的重测序信息显示,等位基因的多样性主要归因于 SSR 区域中重复类型和长度的差异。此外,在微卫星区域的侧翼还观察到一些额外的 INDEL 突变和置换。此外,还发现了一些与防御相关的转录本,如推测的草酸氧化酶(EU024476)和 NBS-LRR 结构域。本研究的EST数据为发现栽培花生的基因和开发SSR标记提供了新的信息来源。共有 16931 个 EST 保存在 NCBI GenBank 数据库中,登录号为 ES751523 至 ES768453。
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引用次数: 0
Wet laboratory tools widely used in plant genomics. 湿式实验室工具广泛应用于植物基因组学。
Pub Date : 2009-01-01 Epub Date: 2010-02-22 DOI: 10.1155/2009/321975
Hikmet Budak, Hongbin Zhang, Pushpendra K Gupta, Boulos Chalhoub, Andrew James, Chunji Liu
The availability of laboratory tools is essential for advanced research in all areas of biological sciences. The recent development of genomic tools has made it possible to deeply investigate and to continuously improve agronomically important traits such as crop yield, quality, and biotic and abiotic stress tolerances. Integrating the newly advanced or developed wet laboratory tools that are widely used in modern genomics research and making them readily accessible will be greatly helpful for research of plant genomics and other disciplines of plant biology. Furthermore, the compilation of the tools will also facilitate scientists to advance the existing tools or develop new tools to address complicated or new questions that were previously intractable in plant genomics and biology. In this special issue of the International Journal of Plant Genomics, “Wet lab tools widely used in plant genomics”, we present the current status of widely used genomics tools and update them with their new advances. Articles published in this special issue cover tools for structural, functional, and comparative genomics and proteomics. The issue also summarizes the advances of genome technology in the past decades and synthesizes the current status of knowledge of new tools with an extension of suggestions. By covering up-to-date genomics tools, this special issue provides a reference for studying the structural and functional organization and evolution of plant genomes. We aim that this special issue will become useful material for teaching and research in plant genomics and biology. There are 8 articles in this special issue, starting with articles on tools for studying plant authopagy (Mitou et al.) and microRNA identification (Unver and Budak), cloning of small RNA (Eric et al.), and the use of virus-induced gene silencing techniques (Unver et al.) for functional analysis of genes and QTLs in plants. Included is also a comprehensive article on heterologous gene expression techniques given by Filiz and Sayers. An up-to-date protocol of Agro-mediated gene transfer in cereal crops is presented by Hensel et al. Additionally, Dechyeva and Schmidt reported one of the critical tools, the molecular cytogenetic mapping of chromosomal fragments and immunostaining of kinetochore proteins, which will greatly help cytogeneticists for identifying and tagging genes in plants, thus promoting plant molecular breeding.
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引用次数: 0
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International journal of plant genomics
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